Antioxidant, Anti-inflammatory or Anti-aging composition containing taxus cambium- or procambium-derived cell line as active ingredient
a technology of anti-inflammatory and anti-aging compositions, applied in hair cosmetics, antinoxious agents, metabolic disorders, etc., can solve the problems of reducing adaptability, reducing metabolic rate, and death of cells and the whole of tissues of the human body
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example 1
Preparation of Taxus Cambium- or Procambium-Derived Cell Line
[0064](1) Preparation of Plant Material
[0065]Each of the twig and stem of Taxus was collected, and then immediately soaked in 100 mg / L of the antioxidant L-ascorbic acid (DUCHEFA, The Netherlands). Then, they were transported and stored.
[0066]Then, the plant was pretreated with a mixed solution of 1% benomyl (Dongbu Hannong Chemical, Korea), 1% daconil (Dongbu Hannong Chemical, Korea), 1% sterptomycin sulphate (DUCHEFA, The Netherlands) and 0.1% cefotaxime sodium (DUCHEFA, The Netherlands) for 24 hours, and then washed with tap water for 30 minutes to remove phenolic compounds and the remaining chemicals. Next, the plant was surface-sterilized in 70% ethanol (DC Chemical, Korea) for 1 min, 30% hydrogen peroxide (LG Chemical, Korea) for 15 min, 1% CLOROX solution for 15 min and 3% CLOROX solution for 5 min, and then washed 3-4 times with water.
[0067](2) Isolation of Procambium and Cambium Tissues from Twig and Stem
[0068]The...
example 2
Preparation of Extract of Procambium- or Cambium-Derived Cell Line
[0081](1) Preparation of DMSO (dimethyl sulfoxide) Extract
[0082](i) 500 g of the cell line from which the medium has been removed were dissolved in 500 ml of DMSO with stirring at 50° C. for 6 hours.
[0083](ii) After completion of the dissolution, the cell solution was centrifuged at 3,000 g for 10 minutes, and the supernatant was collected, thus obtaining a DMSO-soluble substance.
[0084](iii) The obtained DMSO-soluble substance was concentrated using a rotary vacuum concentrator.
[0085](iv) The concentrated sample was dried using a freeze dryer, thereby obtaining a DMSO extract.
[0086](2) Preparation of Distilled Water Extract, Methanol Extract and Acetone Extract
[0087]From the cell line prepared in Example 1, active substances were extracted stepwise as follows (FIG. 2).
[0088](i) 500 g of the cell line from which the culture medium has been removed was dissolved in 500 ml of distilled water with stirring at 50° C. for 6...
example 3
Culture of Human Diploid Fibroblasts (HDF)
[0096]HDF cells were isolated from the fetal penis prepuce and cultured. The culture medium was prepared by adding 10% fetal bovine serum (FBS, Hyclone, Logan, Utah, USA) inactivated by heating at 56° C. for 30 minutes, 100 unit / ml of penicillin, 100 μg g / ml of streptomycin and 300 μg / ml of glutamine to DMEM medium (Invitroge Gibco life tech. Vienna, Austria). The cells were cultured in the medium, described above, in a 5% CO2 incubator at a temperature of 37° C. and a humidity of 95% and subcultured at 3-4-day intervals, immediately before the cells were fused with each other. The subcultured cells were divided, according to the number of subcultures (passages), into young cells cultured less than 20 passages, middle cells cultured for 21-49 passages, and aged cells cultured more than 50 passages. The cultured cells were used in the experiments of Examples 4 to 6.
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